Disease Information: Myobacterium bovis is causing agent for Mycobacterium turberculosis, which is usually transmitted to humans by consuming raw or infected cows milk. It is a chronic bacterial disease of cattle that can affect other species of animals in which evolved from Africa.
Current Inhibitors: Carmustine (BCNU) and other nitrosoureas, Cisplatin, Anthracyclines, Azelaic acid, 13-cis-Retinoic acid, 1-Chloro-2,4-dinitrobenzene, Auranofin and aurothioglucose Expression Information: Expression of wild-type fusion protein and truncated mutant in E. coli Purification Method: recombinant wild-type fusion protein and truncated mutant
TGEADSTDALIGAQR length of your protein in Amino Acids: 710 Molecular Weight: 75723.05 Molar Extinction coefficient of your protein at 280 nm wavelength: 29505 TMpred graph Image
CDS Gene Sequence
<span style="font-family: monospace,serif;"><span class="feature">1..335
/gene="trxB2"
/locus_tag="BQ2027_MB3944"
/coded_by="NC_002945.4:4340100..4341107"
/experiment="experimental evidence, no additional details
recorded"
/note="Mb3944, trxB2, len: 335 aa. Equivalent to Rv3913,
len: 335 aa, from Mycobacterium tuberculosis strain H37Rv,
(99.7% identity in 335 aa overlap). Probable trxB2,
thioredoxin reductase (EC 1.6.4.5) (see citation below),
equivalent to O30973|TRXB_MYCSM THIOREDOXIN REDUCTASE from
Mycobacterium smegmatis (311 aa), FASTA scores: opt: 1575,
E(): 1.8e-87, (78.35% identity in 305 aa overlap); and
highly similar, but shorter at C-terminus, to
P46843|TRXB_MYCLE|TRXB/A|TRX|ML2703 BIFUNCTIONAL
THIOREDOXIN REDUCTASE/THIOREDOXIN from Mycobacterium
leprae (458 aa), FASTA scores: opt: 1766, E(): 8.7e-99,
(83.25% identity in 328 aa overlap). Also highly similar
to many e.g. P52215|TRXB_STRCO|SCH24.12 from Streptomyces
coelicolor (321 aa), FASTA scores: opt: 1249, E():
7.2e-68, (60.4% identity in 313 aa overlap);
Q9Z8M4|TRXB_CHLPN from Chlamydia pneumoniae (Chlamydophila
pneumoniae) (311 aa), FASTA scores: opt: 978, E():
1.3e-51, (49.85% identity in 307 aa overlap);
P09625|TRXB_ECOLI|B0888 from Escherichia coli strain K12
(320 aa), FASTA scores: opt: 948, E(): 8.6e-50, (49.2%
identity in 309 aa overlap); etc. Contains PS00573
Pyridine nucleotide-disulphide oxidoreductases class-II
active site. BELONGS TO THE PYRIDINE NUCLEOTIDE-DISULFIDE
OXIDOREDUCTASES CLASS-II. COFACTOR: FAD (BY SIMILARITY).
Protein product from Mb3944 detected using shotgun mass
spectrometry and SWATH mass spectrometry. Mb3944 found to
be expressed during exponential growth in Sauton's minimal
media by RNA-sequencing."
/transl_table=<span style="color: #642a8f;">[[https://www.ncbi.nlm.nih.gov/Taxonomy/Utils/wprintgc.cgi?mode=c#SG11|11]]</span>
/db_xref="GeneID:<span style="color: #642a8f;">[[https://www.ncbi.nlm.nih.gov/gene/32287584|32287584]]</span>"</span></span>
GC% Content for gene: 10.848401
Primer design results for pNIC-Bsa4 cloning (list seqeunces of all of your ~40 nt long primers): (link to DNA Works output text file - that should be saved in your Google Docs folder after you did the primer design protocol)
Primer design results for 'tail' primers (this is just 2 sequences):
**
Thiredoxin-disulfide Reductase
NCBI Gene 32287584
Protein ID YP_009361307.1
Organism (including strain): Mycobacterium bovis AF2122/97
Etiologic Risk Group:
Disease Information: Myobacterium bovis is causing agent for Mycobacterium turberculosis, which is usually transmitted to humans by consuming raw or infected cows milk. It is a chronic bacterial disease of cattle that can affect other species of animals in which evolved from Africa.
https://www.ncbi.nlm.nih.gov/gene/32287584
Essentiality of this protein:
Is it a monomer or multimer as biological unit? multimer
Complex of proteins?: 2 amino acid chants and 6 ligand in ASU
Druggable Target
https://onlinelibrary.wiley.com/doi/pdf/10.1046/j.1432-1327.2000.01703.x
EC#: 1.8.1.9
Link to BRENDA EC# page:
https://www.brenda-enzymes.org/enzyme.php?ecno=1.8.1.9&Suchword=&reference=&UniProtAcc=&organism%5B%5D=Mycobacterium+tuberculosis
Show screenshot of BRENDA enzyme mechanism schematic
Enzyme Assay information (spectrophotometric, coupled assay ?, reagents):
https://www.sigmaaldrich.com/content/dam/sigma-aldrich/docs/Sigma/Enzyme_Assay/thioredoxinred.pdf
Structure (PDB or Homology model)
PDB #: 2A87
Current Inhibitors: Carmustine (BCNU) and other nitrosoureas, Cisplatin, Anthracyclines, Azelaic acid, 13-cis-Retinoic acid, 1-Chloro-2,4-dinitrobenzene, Auranofin and aurothioglucose
Expression Information: Expression of wild-type fusion protein and truncated mutant in E. coli
Purification Method: recombinant wild-type fusion protein and truncated mutant
Amino Acid Sequence
A87:A|PDBID|CHAIN|SEQUENCE
MTAPPVHDRAHHPVRDVIVIGSGPAGYTAALYAARAQLAPLVFEGTSFGGALMTTTDVENYPGFRNGITGPELMDEMREQ
ALRFGADLRMEDVESVSLHGPLKSVVTADGQTHRARAVILAMGAAARYLQVPGEQELLGRGVSSCATCDGFFFRDQDIAV
IGGGDSAMEEATFLTRFARSVTLVHRRDEFRASKIMLDRARNNDKIRFLTNHTVVAVDGDTTVTGLRVRDTNTGAETTLP
VTGVFVAIGHEPRSGLVREAIDVDPDGYVLVQGRTTSTSLPGVFAAGDLVDRTYRQAVTAAGSGCAAAIDAERWLAEHAA
TGEADSTDALIGAQR
>2A87:B|PDBID|CHAIN|SEQUENCE
MTAPPVHDRAHHPVRDVIVIGSGPAGYTAALYAARAQLAPLVFEGTSFGGALMTTTDVENYPGFRNGITGPELMDEMREQ
ALRFGADLRMEDVESVSLHGPLKSVVTADGQTHRARAVILAMGAAARYLQVPGEQELLGRGVSSCATCDGFFFRDQDIAV
IGGGDSAMEEATFLTRFARSVTLVHRRDEFRASKIMLDRARNNDKIRFLTNHTVVAVDGDTTVTGLRVRDTNTGAETTLP
VTGVFVAIGHEPRSGLVREAIDVDPDGYVLVQGRTTSTSLPGVFAAGDLVDRTYRQAVTAAGSGCAAAIDAERWLAEHAA
TGEADSTDALIGAQR
length of your protein in Amino Acids: 710
Molecular Weight: 75723.05
Molar Extinction coefficient of your protein at 280 nm wavelength: 29505
TMpred graph Image
CDS Gene Sequence
<span style="font-family: monospace,serif;"><span class="feature">1..335 /gene="trxB2" /locus_tag="BQ2027_MB3944" /coded_by="NC_002945.4:4340100..4341107" /experiment="experimental evidence, no additional details recorded" /note="Mb3944, trxB2, len: 335 aa. Equivalent to Rv3913, len: 335 aa, from Mycobacterium tuberculosis strain H37Rv, (99.7% identity in 335 aa overlap). Probable trxB2, thioredoxin reductase (EC 1.6.4.5) (see citation below), equivalent to O30973|TRXB_MYCSM THIOREDOXIN REDUCTASE from Mycobacterium smegmatis (311 aa), FASTA scores: opt: 1575, E(): 1.8e-87, (78.35% identity in 305 aa overlap); and highly similar, but shorter at C-terminus, to P46843|TRXB_MYCLE|TRXB/A|TRX|ML2703 BIFUNCTIONAL THIOREDOXIN REDUCTASE/THIOREDOXIN from Mycobacterium leprae (458 aa), FASTA scores: opt: 1766, E(): 8.7e-99, (83.25% identity in 328 aa overlap). Also highly similar to many e.g. P52215|TRXB_STRCO|SCH24.12 from Streptomyces coelicolor (321 aa), FASTA scores: opt: 1249, E(): 7.2e-68, (60.4% identity in 313 aa overlap); Q9Z8M4|TRXB_CHLPN from Chlamydia pneumoniae (Chlamydophila pneumoniae) (311 aa), FASTA scores: opt: 978, E(): 1.3e-51, (49.85% identity in 307 aa overlap); P09625|TRXB_ECOLI|B0888 from Escherichia coli strain K12 (320 aa), FASTA scores: opt: 948, E(): 8.6e-50, (49.2% identity in 309 aa overlap); etc. Contains PS00573 Pyridine nucleotide-disulphide oxidoreductases class-II active site. BELONGS TO THE PYRIDINE NUCLEOTIDE-DISULFIDE OXIDOREDUCTASES CLASS-II. COFACTOR: FAD (BY SIMILARITY). Protein product from Mb3944 detected using shotgun mass spectrometry and SWATH mass spectrometry. Mb3944 found to be expressed during exponential growth in Sauton's minimal media by RNA-sequencing." /transl_table=<span style="color: #642a8f;">[[https://www.ncbi.nlm.nih.gov/Taxonomy/Utils/wprintgc.cgi?mode=c#SG11|11]]</span> /db_xref="GeneID:<span style="color: #642a8f;">[[https://www.ncbi.nlm.nih.gov/gene/32287584|32287584]]</span>"</span></span>GC% Content for gene: 10.848401Primer design results for pNIC-Bsa4 cloning (list seqeunces of all of your ~40 nt long primers):
(link to DNA Works output text file - that should be saved in your Google Docs folder after you did the primer design protocol)
Primer design results for 'tail' primers (this is just 2 sequences):
**